Purification and properties of F420- and NADP(+)-dependent alcohol dehydrogenases of Methanogenium liminatans and Methanobacterium palustre, specific for secondary alcohols

Bleicher, K.; Winter, J.; Eur. J. Biochem. 200, 43-51 (1991)

Data extracted from this reference:

General Stability

General Stability

Organism

an ammonium sulfate precipitate (3 M, pH 7) of the enzyme loses its activity completely within a few days

Methanobacterium palustre

minor loss of activity in 50 mM phosphate buffer is observed during purification and storage at 4°C without additional precautions

Methanobacterium palustre

no conditions to stabilize the enzyme by freezing at -20°C are found. Addition of 1-10% bovine serum albumin, 20-50% glycerol, 20% 1,2-ethanediol or 0.1% Triton X-100 to the enzyme solution in 40 mM phosphate or 40 mM Tris-HC1 pH 7 before freezing for 1 day or longer has no stabilizing effect

Methanobacterium palustre

Inhibitors

Inhibitors

Commentary

Organism

Structure

1,10-phenanthroline

5 mM, 17% inhibition

Methanobacterium palustre

4 methylpyrazole

2 mM, 17% inhibition

Methanobacterium palustre

4-hydroxymercuribenzoate

0.025 mM, complete inhibition

Methanobacterium palustre

iodoacetate

2 mM, complete inhibition

Methanobacterium palustre

additional information

no inhibition by 5 mM 2,2'-dipyridyl, EDTA does not inhibit up to 5 mM

the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced

724899

Methanobacterium palustre

?

-

-

-

-

additional information

the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced

724899

Methanobacterium palustre DSM 3108

?

-

-

-

-

Subunits

Subunits

Commentary

Organism

tetramer

4 * 44000, SDS-PAGE

Methanobacterium palustre

Temperature Optimum [°C]

Temperature Optimum [°C]

Temperature Optimum Maximum [°C]

Commentary

Organism

37

-

assay at

Methanobacterium palustre

60

-

-

Methanobacterium palustre

Turnover Number [1/s]

Turnover Number Minimum [1/s]

Turnover Number Maximum [1/s]

Substrate

Commentary

Organism

Structure

5

-

2-Pentanol

pH 7.5, 37°C

Methanobacterium palustre

5

-

Cyclohexanol

pH 7.5, 37°C

Methanobacterium palustre

6

-

Cycloheptanone

pH 7.5, 37°C

Methanobacterium palustre

18

-

2-Pentanone

pH 7.5, 37°C

Methanobacterium palustre

26

-

2,3-Butanediol

pH 7.5, 37°C

Methanobacterium palustre

80

-

Cyclopentanone

pH 7.5, 37°C

Methanobacterium palustre

86

-

2-butanone

pH 7.5, 37°C

Methanobacterium palustre

86

-

cyclohexanone

pH 7.5, 37°C

Methanobacterium palustre

106

-

Cyclopentanol

pH 7.5, 37°C

Methanobacterium palustre

123

-

(S)-2-butanol

pH 7.5, 37°C

Methanobacterium palustre

129

-

NADPH

pH 7.5, 37°C

Methanobacterium palustre

129

-

propanone

pH 7.5, 37°C

Methanobacterium palustre

149

-

2-butanol

pH 7.5, 37°C

Methanobacterium palustre

177

-

(R)-2-butanol

pH 7.5, 37°C

Methanobacterium palustre

183

-

acetoin

pH 7.5, 37°C

Methanobacterium palustre

243

-

2-propanol

pH 7.5, 37°C

Methanobacterium palustre

243

-

NADP+

pH 7.5, 37°C

Methanobacterium palustre

609

-

diacetyl

pH 7.5, 37°C

Methanobacterium palustre

pH Optimum

pH Optimum Minimum

pH Optimum Maximum

Commentary

Organism

8

-

-

Methanobacterium palustre

General Stability (protein specific)

General Stability

Organism

an ammonium sulfate precipitate (3 M, pH 7) of the enzyme loses its activity completely within a few days

Methanobacterium palustre

minor loss of activity in 50 mM phosphate buffer is observed during purification and storage at 4°C without additional precautions

Methanobacterium palustre

no conditions to stabilize the enzyme by freezing at -20°C are found. Addition of 1-10% bovine serum albumin, 20-50% glycerol, 20% 1,2-ethanediol or 0.1% Triton X-100 to the enzyme solution in 40 mM phosphate or 40 mM Tris-HC1 pH 7 before freezing for 1 day or longer has no stabilizing effect

Methanobacterium palustre

Inhibitors (protein specific)

Inhibitors

Commentary

Organism

Structure

1,10-phenanthroline

5 mM, 17% inhibition

Methanobacterium palustre

4 methylpyrazole

2 mM, 17% inhibition

Methanobacterium palustre

4-hydroxymercuribenzoate

0.025 mM, complete inhibition

Methanobacterium palustre

iodoacetate

2 mM, complete inhibition

Methanobacterium palustre

additional information

no inhibition by 5 mM 2,2'-dipyridyl, EDTA does not inhibit up to 5 mM

the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced

724899

Methanobacterium palustre

?

-

-

-

-

additional information

the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced